Background: Plant growth is greatly affected by low temperatures, and the expression of a number of genes is induced by cold stress. Although many genes in the cold signaling pathway have been identified in Arabidopsis, little is known about the transcription factors involved in the cold stress response in apple. Results: Here, we show that the apple bHLH (basic helix-loop-helix) gene MdCIbHLH1 (Cold-Induced bHLH1), which encodes an ICE-like protein, was noticeably induced in response to cold stress. The MdCIbHLH1 protein specifically bound to the MYC recognition sequences in the AtCBF3 promoter, and MdCIbHLH1 overexpression enhanced cold tolerance in transgenic Arabidopsis. In addition, the MdCIbHLH1 protein bound to the promoters of MdCBF2 and favorably contributed to cold tolerance in transgenic apple plants by upregulating the expression of MdCBF2 through the CBF (C-repeat-binding factor) pathway. Our findings indicate that MdCIbHLH1 functions in stress tolerance in different species. For example, ectopic MdCIbHLH1 expression conferred enhanced chilling tolerance in transgenic tobacco. Finally, we observed that cold induces the degradation of the MdCIbHLH1 protein in apple and that this degradation was potentially mediated by ubiquitination and sumoylation. Conclusions: Based on these findings, MdCIbHLH1 encodes a transcription factor that is important for the cold tolerance response in apple.

To examine the potential roles of melatonin in cadmium (Cd) uptake, accumulation and detoxification in Malus plants, we exposed two different apple rootstocks varying greatly in Cd uptake and accumulation to either 0 or 30 μM Cd together with 0 or 100 μM melatonin. Cadmium stress stimulated endogenous melatonin production to a greater extent in the Cd-tolerant Malus baccata Borkh. than in the Cd-susceptible Malus micromalus ‘qingzhoulinqin’. Melatonin application attenuated Cd-induced reductions in growth, photosynthesis and enzyme activity, as well as reactive oxygen species (ROS) and malondialdehyde accumulation. Melatonin treatment more effectively restored photosynthesis, photosynthetic pigments and biomass in Cd-challenged M. micromalus ‘qingzhoulinqin’ than in Cd-stressed M. baccata. Exogenous melatonin lowered root Cd²⁺ uptake, reduced leaf Cd accumulation, decreased Cd translocation factors and increased root, stem and leaf melatonin contents in both Cd-exposed rootstocks. Melatonin application increased both antioxidant concentrations and enzyme activities to scavenge Cd-induced ROS. Exogenous melatonin treatment altered the mRNA levels of several genes regulating Cd uptake, transport and detoxification including HA7, NRAMP1, NRAMP3, HMA4, PCR2, NAS1, MT2, ABCC1 and MHX. Taken together, these results suggest that exogenous melatonin reduced aerial parts Cd accumulation and mitigated Cd toxicity in Malus plants, probably due to the melatonin-mediated Cd allocation in tissues, and induction of antioxidant defense system and transcriptionally regulated key genes involved in detoxification.

Non-bagging apple cultivation, which is time-saving, labor-saving, and cost-effective, represents the future direction of apple cultivation in China. However, compared with bagging cultivation, it degrades fruit appearance quality, characterized by rough peels and dull colors, with the underlying physiological and molecular mechanisms remaining unclear. This study used ‘Tianhong 2’ Fuji apples, grafted onto SH dwarfing rootstock, and integrated transcriptomics–metabolomics to explore these mechanisms. Results showed that non-bagging-cultivated apple peels had higher chlorophyll and carotenoid contents but lower anthocyanin content than those of bagging-cultivated ones. Transcriptome sequencing identified 1571 differentially expressed genes (DEGs: 1269 upregulated, 302 downregulated). Functional analysis revealed that the decline in fruit appearance quality was primarily associated with secondary metabolite biosynthesis, lipid metabolism, and carbohydrate metabolism, and 34 candidate genes were identified. Metabolomic analysis detected 394 differentially expressed metabolites (DEMs), 38 of which were closely related to the non-bagging-induced appearance degradation, mainly lipids, organic oxygen compounds, and organic acids and their derivatives. Integrated analysis of DEGs and DEMs indicated the involvement of multiple critical metabolic pathways, including cutin, suberin and wax biosynthesis; starch and sucrose metabolism; cyanoamino acid metabolism; and phenylpropanoid and flavonoid biosynthesis. Compared with bagging-cultivated apples, non-bagging-cultivated apples exhibited faster starch degradation and higher soluble sugar accumulation. Additionally, the accumulation of specific metabolites [e.g., quercetin (HMDB0005794, HMDB03249, LMPK12112097), and suberin components (LMFA01170020, LMFA01050437, HMDB0031885)], along with elevated organic acid levels, contributed to peel roughness and dull coloration. These findings further enrich the theoretical basis for the formation of fruit quality in Fuji apples under non-bagging cultivation and provide valuable theoretical guidance for the practical application of this cultivation mode.

Citrus leaf blotch virus (CLBV, genus Citrivirus, Family Betaflexiviridae) is often associated with yellow vein clearing, leaf mottle, and bud union crease symptoms on infected plants. CLBV is repo...

Improving apple peel color has been an important objective in apple production. To better understand the effect and mechanism of the combined application of urea and FA (fulvic acid) regulation of anthocyanin biosynthesis, a field experiment was performed in 2022 and 2023, respectively, under five treatments of urea + FA (CK, urea only; FA50, urea + 50 kg ha⁻¹ FA; FA100, urea + 100 kg ha⁻¹ FA; FA150, urea + 150 kg ha⁻¹ FA; FA200, urea + 200 kg ha⁻¹ FA), using isotope (¹³C and ¹⁵N) marking to analyze the changes in carbon (C) and nitrogen (N) nutrient distribution as well as anthocyanin biosynthesis in fruits. We observed that, under FA application conditions, anthocyanin content in the peel was elevated in both years, with increases of 15.98~52.88% in 2022 and 15.93~52.94% in 2023. The best promotion effects were observed under FA150 treatment. Apart from the expression levels of anthocyanin biosynthesis-related genes and transcription factors in the apple peel, this positive effect on anthocyanin content induced by FA addition was also found to be associated with the optimization of C and N distribution in leaves and fruits. On the one hand, the application of FA not only enhanced leaf photosynthetic-related indexes (such as P_n, G_s, and Rubisco activity) and influenced (increased) S6PDH, SPS, and SS activities in leaves, but also elevated fruit sugar metabolism-related enzyme (SDH, SS-c, AI, and NI) activity and upregulated fruit stalk sugar transporter (MdSOT1, MdSOT3, MdSUT1 and MdSUT4) gene expression, which ultimately promoted the synthesis and the leaf to fruit transport of photosynthates, thus promoting ¹³C-photosynthate accumulation in fruits. On the other hand, FA application elevated leaves’ N metabolism-related enzyme (GS and GOGAT) activity and optimized ¹⁵N distribution in leaves and fruits. Moreover, we also observed that FA application altered the fate of N fertilizer in apple orchards, showed an elevation in apple tree ¹⁵NUE and soil ¹⁵N residuals and showed a decrease in soil ¹⁵N loss. In summary, the appropriate application of FA150 (urea + 150 kg ha⁻¹) synergistically optimized C and N nutrient distribution, and promoted anthocyanin biosynthesis in apple trees.